Osteogenic differentiation: MSCs grown on 24 wells plate (TPP, Switzerland) for up to 80% of confluence. Then growth media was replaced with StemPro™ Osteogenesis Differentiation media (Gibco, Thermo Fisher Scientific, USA). MSCs were refeed every 3–4 days. After 21 days of osteogenic differentiation induction MSCs were stained with Alizarin Red S to stain calcium deposits, marker of mature osteoblasts [38 (link)].
Chondrogenic differentiation: MSCs grown on 24 wells plate (TPP, Switzerland) for up to 80% of confluence. Then growth media was replaced with MSCgo™ Chondrogenic Differentiation Medium (Sartorius, Germany). MSCs were refeed every 3–4 days. After 21 days of chondrogenic differentiation induction MSCs were stained with Alcian Blue to stain acidic polysaccharides [39 (link)].
Adipogenic differentiation: MSCs grown on 24 wells plate (TPP, Switzerland) for up to 80% of confluence. Then growth media was replaced with MesenCult™ Adipogenic Differentiation Kit (Human) (STEMCELL Technologies Inc., Canada). MSCs were refeed every 3–4 days. After 21 days of adipogenic differentiation induction MSCs were stained with Oil Red O to lipid drops [40 (link)].
Chondrogenic differentiation: MSCs grown on 24 wells plate (TPP, Switzerland) for up to 80% of confluence. Then growth media was replaced with MSCgo™ Chondrogenic Differentiation Medium (Sartorius, Germany). MSCs were refeed every 3–4 days. After 21 days of chondrogenic differentiation induction MSCs were stained with Alcian Blue to stain acidic polysaccharides [39 (link)].
Adipogenic differentiation: MSCs grown on 24 wells plate (TPP, Switzerland) for up to 80% of confluence. Then growth media was replaced with MesenCult™ Adipogenic Differentiation Kit (Human) (STEMCELL Technologies Inc., Canada). MSCs were refeed every 3–4 days. After 21 days of adipogenic differentiation induction MSCs were stained with Oil Red O to lipid drops [40 (link)].
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